Flow Cytometry,HPLC-RP,and metAFLP Analyses to Assess Genetic Variability in Somatic Embryo-Derived Plantlets of <Emphasis Type="BoldItalic">Gentiana pannonica</Emphasis> Scop |
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Authors: | Agnieszka Fiuk Piotr T Bednarek Jan J Rybczyński |
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Institution: | 1.Botanical Garden–Center for Biological Diversity Conservation,Polish Academy of Sciences,Warsaw,Poland;2.Department of Plant Biochemistry and Physiology,Plant Breeding and Acclimatization Institute,B?onie,Poland |
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Abstract: | Cytometric and molecular techniques were used to verify genetic uniformity among somatic embryo-derived plantlets of Gentiana pannonica Scop. Cytometric analysis of regenerants revealed absence of chromosomal changes and alterations in ploidy. However, reverse
phase high pressure liquid chromatography detected higher levels of methylation in regenerated plants than those of control
plants. These changes were further investigated using a quantitative molecular marker-based approach. This revealed that numerous
tissue culture-induced variations, ∼3% (epi)mutations, were observed, including sequence variation and changes in methylation
patterns. Moreover, complex patterns of variation, including combinations of genetic and epigenetic changes, were relatively
high (ca. 9%). Overall, tissue culture-induced variation reached 16%; while, demethylation was lower than de novo methylation
in heterozygotic material and similar in all regenerated plantlets. |
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